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Related Concept Videos

CRISPR01:59

CRISPR

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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RNA Polymerase II Accessory Proteins02:36

RNA Polymerase II Accessory Proteins

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Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
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Related Experiment Video

Updated: Feb 28, 2026

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
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Repurposing CRISPR System for Transcriptional Activation.

Meng Chen1,2, Lei Stanley Qi3,4,5

  • 1Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.

Advances in Experimental Medicine and Biology
|June 23, 2017
PubMed
Summary

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, when modified to a deactivated form (dCas9), enables precise gene activation. This CRISPR/dCas9 technology offers a powerful and scalable method for endogenous gene regulation.

Keywords:
CRISPR/CasEndogenous gene activationGene regulationTranscriptional activation

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Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a leading genome editing tool.
  • Mutating Cas9's nuclease domains yields a deactivated form (dCas9) that binds DNA without cleaving it.
  • This dCas9 retains RNA-guided DNA-targeting capabilities, enabling applications beyond gene editing.

Purpose of the Study:

  • To review recent literature on CRISPR/dCas9-based transcriptional activation systems.
  • To highlight the versatility of CRISPR/dCas9 for genome imaging, gene regulation, and epigenetic modification.
  • To compare CRISPR/dCas9-mediated gene activation with conventional methods.

Main Methods:

  • Utilizing deactivated Cas9 (dCas9) fused with activation domains.
  • Employing RNA-guided targeting for specific DNA sequence binding.
  • Genome-wide transcriptional activation across diverse organisms.

Main Results:

  • CRISPR/dCas9 systems facilitate specific and efficient transcriptional activation.
  • Fusion of dCas9 with activation domains enables scalable gene expression enhancement.
  • CRISPR/dCas9 technology demonstrates robustness and convenience for gene regulation.

Conclusions:

  • CRISPR/dCas9-based transcriptional activation is a promising technology for genome regulation.
  • This system offers specificity, convenience, robustness, and scalability for endogenous gene activation.
  • CRISPR/dCas9 represents an advancement over conventional gene expression enhancement approaches.